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Retina: ultrastructural alterations produced by extremely low levels of coherent radiation.

The effect of low levels of coherent radiation on the eye is not fully established, but is generally presumed to be noninjurious. Irradiation of the retina with a Q-switched ruby laser emitting low amounts of energy (0.1 percent probability of creating damage) consistently produces ultrastructural alterations in rods and cones. Outer segments of these cells are broken and disorganized and their lamellae are in disarray 1 day after such irradiation.

Animals↗

Novel tellurite-amended media and specific chromosomal and Ti plasmid probes for direct analysis of soil populations of Agrobacterium biovars 1 and 2.

Ecology and biodiversity studies of Agrobacterium spp. require tools such as selective media and DNA probes. Tellurite was tested as a selective agent and a supplement of previously described media for agrobacteria. The known biodiversity within the genus was taken into account when the selectivity of K(2)TeO(3) was analyzed and its potential for isolating Agrobacterium spp. directly from soil was evaluated. A K(2)TeO(3) concentration of 60 ppm was found to favor the growth of agrobacteria and restrict the development of other bacteria. Morphotypic analyses were used to define agrobacterial colony types, which were readily distinguished from other colonies. The typical agrobacterial morphotype allowed direct determination of the densities of agrobacterial populations from various environments on K(2)TeO(3)-amended medium. The bona fide agrobacterium colonies growing on media amended with K(2)TeO(3) were confirmed to be Agrobacterium colonies by using 16S ribosomal DNA (rDNA) probes. Specific 16S rDNA probes were designed for Agrobacterium biovar 1 and related species (Agrobacterium rubi and Agrobacterium fici) and for Agrobacterium biovar 2. Specific pathogenic probes from different Ti plasmid regions were used to determine the pathogenic status of agrobacterial colonies. Various morphotype colonies from bulk soil suspensions were characterized by colony blot hybridization with 16S rDNA and pathogenic probes. All the Agrobacterium-like colonies obtained from soil suspensions on amended media were found to be bona fide agrobacteria. Direct colony counting of agrobacterial populations could be done. We found 10(3) to 10(4) agrobacteria. g of dry soil(-1) in a silt loam bulk soil cultivated with maize. All of the strains isolated were nonpathogenic bona fide Agrobacterium biovar 1 strains.

Colony Count, Microbial↗

Diversity of bacteria associated with natural aphid populations.

The bacterial communities of aphids were investigated by terminal restriction fragment length polymorphism and denaturing gradient gel electrophoresis analysis of 16S rRNA gene fragments generated by PCR with general eubacterial primers. By both methods, the gamma-proteobacterium Buchnera was detected in laboratory cultures of six parthenogenetic lines of the pea aphid Acyrthosiphon pisum and one line of the black bean aphid Aphis fabae, and one or more of four previously described bacterial taxa were also detected in all aphid lines except one of A. pisum. These latter bacteria, collectively known as secondary symbionts or accessory bacteria, comprised three taxa of gamma-proteobacteria (R-type [PASS], T-type [PABS], and U-type [PAUS]) and a rickettsia (S-type [PAR]). Complementary analysis of aphids from natural populations of four aphid species (A. pisum [n = 74], Amphorophora rubi [n = 109], Aphis sarothamni [n = 42], and Microlophium carnosum [n = 101]) from a single geographical location revealed Buchnera and up to three taxa of accessory bacteria, but no other bacterial taxa, in each aphid. The prevalence of accessory bacterial taxa varied significantly among aphid species but not with the sampling month (between June and August 2000). These results indicate that the accessory bacterial taxa are distributed across multiple aphid species, although with variable prevalence, and that laboratory culture does not generally result in a shift in the bacterial community in aphids. Both the transmission patterns of the accessory bacteria between individual aphids and their impact on aphid fitness are suggested to influence the prevalence of accessory bacterial taxa in natural aphid populations.

Animals↗

Rhizobial 16S rRNA and dnaK genes: mosaicism and the uncertain phylogenetic placement of Rhizobium galegae.

The phylogenetic relatedness among 12 agriculturally important species in the order Rhizobiales was estimated by comparative 16S rRNA and dnaK sequence analyses. Two groups of related species were identified by neighbor-joining and maximum-parsimony analysis. One group consisted of Mesorhizobium loti and Mesorhizobium ciceri, and the other group consisted of Agrobacterium rhizogenes, Rhizobium tropici, Rhizobium etli, and Rhizobium leguminosarum. Although bootstrap support for the placement of the remaining six species varied, A. tumefaciens, Agrobacterium rubi, and Agrobacterium vitis were consistently associated in the same subcluster. The three other species included Rhizobium galegae, Sinorhizobium meliloti, and Brucella ovis. Among these, the placement of R. galegae was the least consistent, in that it was placed flanking the A. rhizogenes-Rhizobium cluster in the dnaK nucleotide sequence trees, while it was placed with the other three Agrobacterium species in the 16S rRNA and the DnaK amino acid trees. In an effort to explain the inconsistent placement of R. galegae, we examined polymorphic site distribution patterns among the various species. Localized runs of nucleotide sequence similarity were evident between R. galegae and certain other species, suggesting that the R. galegae genes are chimeric. These results provide a tenable explanation for the weak statistical support often associated with the phylogenetic placement of R. galegae, and they also illustrate a potential pitfall in the use of partial sequences for species identification.

Alleles↗

Unconventional genomic organization in the alpha subgroup of the Proteobacteria.

Pulsed-field gel electrophoresis was used to analyze the genomic organization of 16 bacteria belonging or related to the family Rhizobiaceae of the alpha subgroup of the class Proteobacteria. The number and sizes of replicons were determined by separating nondigested DNA. Hybridization of an rrn gene probe was used to distinguish between chromosomes and plasmids. Members of the genus Agrobacterium all possess two chromosomes, and each biovar has a specific genome size. As previously demonstrated for Agrobacterium tumefaciens C58, the smaller chromosomes of Agrobacterium biovar 1 and Agrobacterium rubi strains appear to be linear. The genomes of Rhizobium strains were all of similar sizes but were seen to contain either one, two, or three megareplicons. Only one chromosome was present in the member of the related genus Phyllobacterium. We found one or two chromosomes in Rhodobacter and Brucella species, two chromosomes in Ochrobactrum anthropi, and one chromosome in Mycoplana dimorpha and Bartonella quintana; all of these genera are related to the Rhizobiaceae. The presence of multiple chromosomes is discussed from a phylogenetic and taxonomic point of view.

Bartonella↗

COMPARATIVE PHYSIOLOGY OF ANTIBIOTIC-RESISTANT STRAINS OF STAPHYLOCOCCUS AUREUS.

Brown, Ruby L. (North Carolina State College, Raleigh) and James B. Evans. Comparative physiology of antibiotic-resistant strains of Staphylococcus aureus. J. Bacteriol. 85:1409-1412. 1963.-A collection of antibiotic-resistant strains of Staphylococcus aureus isolated from clinical sources was studied with respect to nutritional requirements and common diagnostic tests. Contrary to numerous reports in the literature indicating changes in these characteristics in antibiotic-resistant mutants, the present cultures were typical members of the taxonomic species S. aureus. They were coagulase-positive, fermented both glucose and mannitol under anaerobic conditions, produced acetoin from glucose, grew and produced black colonies on tellurite glycine agar, required both thiamine and nicotinic acid, and did not require other vitamins or purines. It is suggested that in most instances these cultures from clinical sources represent spontaneous mutants having genetic changes limited largely to loci concerned with antibiotic resistance. Most reports of extensive changes in physiology and nutritive requirements by antibiotic-resistant strains of S. aureus are based on studies of resistant strains selected after exposing a large population of the parent sensitive strain to toxic levels of antibiotics, chemical mutagens, or irradiation. Such isolates may have widespread genetic damage at other loci in addition to those concerned with their antibiotic resistance.

Anti-Bacterial Agents↗

Comparative study of surgical instruments from sterile-service departments for presence of residual gram-negative endotoxin and proteinaceous deposits.

The ineffective cleaning of surgical instruments may be a vector for the transmission of hospital-acquired infections. The aim of this research was to investigate whether further decontamination procedures need to be instigated in sterile-service departments (SSDs) to reduce the risk of nosocomial illnesses, such as endotoxemia, sepsis, or iatrogenic Creutzfeldt-Jakob disease (to date, 1,147 cases of confirmed Creutzfeldt-Jakob disease deaths in the United Kingdom since 1990 have been reported). Instrument sets were obtained from nine anonymous United Kingdom National Health Service (NHS) primary care trust SSDs. The investigation implemented an advanced light microscopy technique, episcopic differential interference contrast microscopy with the sensitive fluorescent reagents SYPRO Ruby and 4',6-diamidino-2-phenylindole dihydrochloride (DAPI), to detect proteinaceous and microbial contamination levels. Gram-negative lipopolysaccharide (LPS) endotoxin was monitored using a dansylated polymyxin B fluorochrome agent. None of the 260 instruments examined displayed signs of microbial colonization or LPS endotoxin contamination. However, over 60 percent of the instruments showed a high degree of protein soiling (0.4 to 4.2 mug protein/mm(2)). Some instruments appeared soiled with crystalline deposits that may consist of a potentially hazardous material contributing to inflammation and/or surgical shock. It is clear that the overall standard for cleaning must be raised in order to fulfill the imminent introduction of new European standards and to reduce the risk of cross-patient contamination and iatrogenic transmission.

Decontamination↗

Response of the human eye to laser irradiation of the iris.

Irradiation of the iris of glaucoma patients with either a ruby or an argon laser caused no obvious disruption of the blood-aqueous barrier (as visualised by fluorescein angiography), in contrast to a sudden and pronounced effect in the rabbit eye. Only a low and variable increase in intraocular pressure occurred in the human subjects after the laser irradiation in comparison with a consistently high rise of ocular tension in the rabbit. This investigation indicates that the human eye is much less responsive to injury.

Aged↗

Neodymium-YAG transscleral cyclocoagulation in rabbit eyes.

Transscleral ruby cyclocoagulation has been successfully used to lower intraocular pressure. The recent commercial availability of Nd-YAG lasers with a thermal mode provides a possible alternative by which to perform this procedure. We treated one eye of seven pigmented rabbits using the thermal mode of the Nd-YAG laser. The intraocular pressures were followed up for three months. The treated eyes had significantly lower mean intraocular pressures were followed up for three months. The treated eyes had significantly lower mean intraocular pressures than the untreated contralateral eyes (p less than 0.001). Moreover, the decrease was sustained over the three-month duration of the study. Pathological examination revealed very selective destruction of the ciliary processes, with sparing of the overlying ciliary muscle, sclera, and conjunctiva. The potential value of this mode of therapy for use in patients with glaucoma is discussed.

Animals↗

Laser iridotomy.

A ruby laser has been used to produce a permeable lesion in the iris to establish a communication between the anterior and posterior chambers. In a preliminary study in nine patients the technique gave satisfactory results in the prophylactic treatment of four cases of incipient closed-angle glaucoma and of two cases of iris bombé following uveitis.

Anterior Chamber↗

Counteraction of platelet activity at sites of laser-induced endothelial trauma.

The effects of dextran 70 (Mw 70,000; Macrodex), heparin, dipyridamole (Persantin), and prostaglandin E(1) on the behaviour of platelets were investigated at sites of endothelial trauma induced with a ruby biolaser in arterioles in the rabbit ear chamber. This technique has several advantages over previous methods of studying platelet activity.Dextran 70 (2 g./kg. body weight) caused a profound reduction in platelet activity four to six hours after its administration. Heparin (12 mg./kg. body weight) had no effect. The effect of dipyridamole (2.5 mg./kg. body weight) was profound immediately after its administration, but was transient. In preliminary experiments single intravenous injections of prostaglandin E(1) (25 and 125 mug./kg. body weight) had no detectable effect.

Animals↗

Dominant-and-recessive epistasis in a homeotic mosquito mutant.

Following selection for 15 generations a pure strain of a homeotic mutant spur was isolated from a Brazilian population of the mosquito Culex pipiens fatigans. Monohybrid crosses showed a 13:3 segregation indicating dominant-and-recessive epistasis for wild-type vs. spur. This implies that a dominant allele at one locus and a recessive at the other interact to produce the mutant phenotype. Dihybrid crosses with linkage group II markers yellow and ruby gave 39:13:9:3 ratios indicating independent segregation. However, the dihybrid cross with linkage group I marker maroon showed a highly significant departure from 39:13:9:3 ratio. Data available indicate that the phenotype spur is controlled by a dominant epistat in linkage group III and a recessive epistat (approximately 31.9 crossover units from maroon) in linkage group I.

Alleles↗

Role of glycogen synthase kinase-3 in TNF-alpha-induced NF-kappaB activation and apoptosis in hepatocytes.

Nuclear factor-kappaB (NF-kappaB) prevents hepatocytes from undergoing apoptosis during development and liver regeneration. Mice with inactivated glycogen synthase kinase (GSK)-3beta die from hepatocyte apoptosis during development due to a defect in NF-kappaB activation (Hoeflich KP, Luo J, Rubie EA, Tsao MS, Jin O, and Woodgett JR. Nature 406: 86-90, 2000). In this study, we determined the role of GSK-3 in TNF-alpha-induced NF-kappaB activation and cell death in primary hepatocytes. LiCl, an established inhibitor of GSK-3, sensitized primary rat hepatocytes toward TNF-alpha-mediated apoptosis resulting in 90% cell death after 24 h. This was accompanied by increased caspase 8-like and 3-like activities, nuclear fragmentation and DNA laddering. LiCl treatment had no effect on IkappaB-alpha degradation, IkappaB kinase (IKK) activity, NF-kappaB binding activity, and p65 nuclear import and export, but decreased transcription of the NF-kappaB-dependent inducible nitric oxide synthase gene and a NF-kappaB-driven reporter gene. The p65 sequence revealed four potential GSK-3 phosphorylation sites within its COOH-terminal transactivation domains and recombinant GSK-3beta phosphorylated glutathione S-transferase (GST)-p65(354-551), but not GST-p65(1-305) in vitro. These results indicate that GSK-3 protects hepatocytes from TNF-alpha-induced apoptosis through p65 phosphorylation and upregulation of NF-kappaB transactivation.

Animals↗

Gastrointestinal tract innervation of the mouse: afferent regeneration and meal patterning after vagotomy.

Mice, with the variety of genotypes they provide, should be particularly useful for studies of growth factors and gene products in regeneration of autonomic pathways such as the vagus nerve. To provide a foundation for examinations of mouse vagal reorganization, two experiments assessed the rate, extent, and accuracy of afferent reinnervation of the stomach after vagotomy and related these patterns to feeding behavior. In experiment 1, the pattern of afferent regrowth into the gut after unilateral truncal vagotomy was characterized by labeling of these afferents with wheat germ agglutinin-horseradish peroxidase and Micro-Ruby. Regenerating neurites had reached and, in some cases, already reinnervated the stomach by 4 wk after axotomy. By 8 wk, regrowth was more extensive, and many fibers had redifferentiated terminals in the smooth muscle. By 16 wk, vagal projections had reached or exceeded normal density in the corpus, density in the forestomach was still reduced, and regrowth in the antrum was minimal. At all time points, not only appropriate terminals, but also growth cones and aberrant endings, were observed. In experiment 2, meal patterns of vagotomized mice were evaluated using a solid diet over the period of regeneration; cholecystokinin suppression of a liquid meal after unilateral and bilateral truncal vagotomies was also evaluated. Unilaterally, as well as bilaterally, vagotomized animals ate smaller and more frequent meals. These disturbed patterns became more pronounced in the first 8 wk after vagotomy, during regeneration. Cholecystokinin inhibition of intake was attenuated by bilateral, but not unilateral, vagotomy. Overall, the spatial and temporal patterns of structural and functional changes observed during regeneration verify that the mouse provides a useful preparation for examining the control of vagal plasticity.

Afferent Pathways↗

Sodium loading changes urinary protein excretion: a proteomic analysis.

Plasma sodium concentration is maintained even when sodium intake is altered. Sodium homeostasis may involve changes in renal tubular protein expression that are reflected in the urine. We used proteomic analysis to investigate changes in urinary protein excretion in response to acute sodium loading. Rats were given deionized water followed by hypertonic (2.7%) saline for 28 h each. Urinary protein expression was determined during the final 4 h of each treatment. Acute sodium loading increased urinary sodium excretion (4.53 +/- 1.74 vs. 1.70 +/- 0.27 mmol/day, P = 0.029). Urinary proteins were separated by two-dimensional PAGE and visualized by Sypro ruby staining. Differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry followed by peptide mass fingerprinting. The abundance of a total of 45 protein components was changed after acute sodium loading. Neutral endopeptidase, solute carrier family 3, meprin 1alpha, diphor-1, chaperone heat shock protein 72, vacuolar H(+)-ATPase, ezrin, ezrin/radixin/moesin-binding protein, glutamine synthetase, guanine nucleotide-binding protein, Rho GDI-1, and chloride intracellular channel protein 1 were decreased, whereas albumin and alpha-2u globulin were increased. Some of these proteins have previously been shown to be associated with tubular transport. These data indicate that alterations in the excretion of several urinary proteins occur during acute sodium loading.

Amino Acid Sequence↗

Projections from primary somatosensory cortex to the neostriatum: the role of somatotopic continuity in corticostriatal convergence.

We characterized the organization of corticostriatal projections from rodent primary somatosensory cortex (SI), testing the hypothesis that projections from SI areas representing subcomponents of the forelimb exhibit greater neostriatal overlap than projections from areas representing separate body parts. The anterograde tracers Fluoro-Ruby (FR), Alexa Fluor (AF), and biotinylated dextran amine (BDA) were injected into physiologically identified regions of rat SI. Injection locations were confirmed by examining the SI barrel fields and limb representations in tangential sections processed for cytochrome oxidase (CO). Experimental animals were divided into two groups: one group received multiple tracer injections in neighboring SI regions that represent separate body parts (whiskers, forepaw, and hindpaw); the other group received injections in SI areas that represent different components of the forelimb (forepaw, antebrachium, and brachium). The distribution of labeled terminals and their varicosities in the neostriatum and in the thalamus were plotted and quantitatively analyzed. For most animals, tracer overlap in the thalamus was either minimal or completely absent. In the neostriatum, projections from the whisker, forelimb, and hindlimb representations terminated in regions that rarely overlap with each other, while those originating from different parts of the forelimb representation were more likely to terminate in overlapping parts of the neostriatum. To the extent that neostriatal activation depends on corticostriatal convergence, the corticostriatal projections in the sensorimotor channel appeared to be organized so that neostriatal neurons may signal when multiple components of the same body part are activated simultaneously.

Animals↗

Role of laser therapy in dermatology -- clinical aspects.

BACKGROUND: Laser therapy has become an integral part of therapeutic alternatives in dermatology. Nevertheless, there are only few statistical data on the role of lasers in everyday practice of dermatological clinics. OBJECTIVE: Analysis of laser patients, indications and lasers used. METHODS: We analysed 10244 patients treated with laser therapy in the Department of Dermatology at the University of Regensburg from 1992 to 2000. This figure represents 9.6 % of all patients seen during that period. RESULTS: Vascular lesions (hemangiomas, port-wine stains, telangiectasias) represented the most common indication (more than 70%) followed by infectious disorders (mostly papillomavirus lesions) as well as the removal of hairs and tattoos. Accordingly, the most frequently used laser was the flashlamp-pumped pulsed dye laser (585 nm) followed by the argon ion, the long-pulse tunable flashlamp-pumped pulsed dye laser, the Q-switch ruby laser, and the CO(2)-laser. In recent years, the percentage of indications and lasers used has changed significantly, reflecting the development of new lasers and the introduction of new indications. CONCLUSIONS: The results confirm the important role of laser therapy in dermatology.

Adolescent↗